Table 1.
Substrates, adaptors, and modifiers/regulators of NEDD4-1.
| Name | Substrate | Adaptor | Modifier/Regulator | Binding Motifs | Modification | Function | Experimental Model Used | References |
|---|---|---|---|---|---|---|---|---|
| ABCB1 (ATP-binding cassette sub-family B member 1/P-Glycoprotein) | 🗸 | PDY | Polyubiquitination | An ATP Binding Cassette transporter that exports β-amyloid from Blood-brain barrier endothelial cells. Potential for intervention in Alzheimer’s disease | In vitro: Sf21 cells | [31] | ||
| α-synuclein | 🗸 | PDNEAYEMP, PLY, PPLP, PPSP, PFY | Monoubiquitination and Polyubiquitination (K63–linked) | Lysosomal degradation. Potential protection mechanism against Parkinson’s Disease pathogenesis | In vitro: SH–SY5Y cells In vivo: Drosophila and Rat |
[28,29,30] | ||
| Beclin 1 | 🗸 | LPLY | Beclin 1: polyubiquitination (K63 and K11–linked) | Subunit of the class III phosphatidylinositol 3-kinase complex. Autophagy-related protein. Proteasomal degradation. Inhibited autophagy and cell survival | In vitro: HeLa cells | [32] | ||
| β2-AR (β2-Arrestin Receptor) | 🗸 | Ubiquitination | Involved in internalised receptor degradation and lysosomal trafficking. Degraded via the lysosome | In vitro: HEK293 cells | [33] | |||
| β-arrestin 1 | 🗸 | Adaptor protein for NHE1 ubiquitination | In vitro: HEK293 cells, mouse embryonic fibroblasts | [34] | ||||
| β-arrestin 2 | 🗸 | 🗸 | Adaptor protein for β2-adrenergic receptor ubiquitination | In vitro: HEK293 cells | [33] | |||
| Caspase-1, -3, -4, -6, -7, -11 | 🗸 | 🗸 | Truncation | NEDD4-1 can be cleaved by caspases and can K48-polyubiquitinate caspase 11 | In vitro: Jurkat cells A549 | [14,35,36,37] | ||
| Cbl-b (Casitas B-lineage Lymphoma b) | 🗸 | 🗸 | Polyubiquitination | Impedes NEDD4-1 interaction with PTEN and also polyubiquitinates Cbl-b for degradation | In vivo: Cbl-bC373A and NEDD4Gt(IRESβgeo)249Lex mice | [38,39] | ||
| CNrasGEF (Cyclic Nucleotide rat sarcoma virus Guanine nucleotide Exchange Factor) | 🗸 | PPGY, PPDY | Polyubiquitination | RAS guanine nucleotide exchange factors that are degraded via the proteasome. NEDD4-1 overexpression promoted migration and invasion of glioma cells | In vitro: HEK293T and Glioma cells | [40,41] | ||
| Connexin43 | 🗸 | PPGY | Ubiquitination | Gap junction protein. Proteasome and lysosome degradation | In vitro: WB-F344 rat liver epithelial cells | [42] | ||
| c-Src (Proto-oncogene tyrosine-protein kinase Src) | 🗸 | Tyrosine kinase that activates NEDD4-1 through phosphorylation of its HECT and C2 domains. Phosphorylation inhibits auto-regulation | In vitro: HeLa, HEK293T and Platinum E cells | [43] | ||||
| Δ Np63 transcriptional target | 🗸 | 🗸 | PPPY | Ubiquitination and polyubiquitination | A homologue of p53 tumour suppressor. Protein destabilisation of ∆Np63α and degradation. Downregulates NEDD4-1 leading to the suppression of nuclear PTEN in basal layer keratinocytes |
In vitro: HEK293-EBNA, HaCaT, A431 andH1299 cells In vivo: Zebrafish embryos | [44,45] | |
| EPS15 (Epithelial growth factor receptor substrate 15) |
🗸 | Monoubiquitination and polyubiquitination | An endocytic protein that is targeted for degradation by NEDD4-1 | In vitro: HeLa and B82L cells | [46] | |||
| FGFR1 (Fibroblast Growth Factor Receptor 1) | 🗸 | 🗸 | VLLVRPSRLSSSG | Ubiquitination | FGFR1 is a tyrosine kinase involved in cell proliferation and differentiation during development. Inhibited neural stem cell differentiation. Activates c-Src that subsequently activates NEDD4-1 | In vitro: Human embryonic stem cells In vivo: Zebrafish embryos |
[47,48] | |
| GAG (group specific antigen) protein | 🗸 | PPPY, PTAP | Monoubiquitination | From the HTLV-1 (human T-lymphotropic virus type 1) involved in hijacking mutivesicular body (MVB) pathway proteins required for viral budding | In vitro: HEK293T cells | [49] | ||
| γ2-adaptin | 🗸 | 🗸 | PPAY | Monoubiquitination and multi-polyubiquitination | A member of the clathrin adaptor protein family. Forms a complex with NEDD4-1 and is involved in endosomal/multivesicular body (MVB) pathway and the assembly and release of the HBV | In vitro: HuH-7 and HeLa cells | [50,51] | |
| HBV X protein (Hepatitis B virus) | 🗸 | Ubiquitination | A multifunctional regulator that is encoded by the HBV genome. It is degraded via the K48 proteasomal pathway | In vitro: HEK293T, HBV-related HCC cell lines - HepG2.215, HepG3B, SNU182, SNU387, PLC/PRF/5, and MHCC97H | [52] | |||
| HER3 (human epidermal growth factor receptor 3) | 🗸 | PPRY | Polyubiquitination | A member of the EGRF family. Degraded via the proteasome. Inhibited cancer cell proliferation and tumour growth. NEDD4-1 knockdown induces apoptosis in DU145 cells | In vivo: Chinese Hamster ovary cells In vitro: MDA-MB-453, MCF-7, and DU145 cells |
[53,54] | ||
| HGS (Hepatocyte growth factor-regulated tyrosine kinase substrate) | 🗸 | PPEY | Ubiquitination | Binding partner for NEDD4-1 involved in EGFR lysosome degradation | In vitro: Chinese hamster ovary and HeLa cells | [55] | ||
| IFITM3 (Interferon (IFN)-induced transmembrane protein 3) | 🗸 | PPNY | Polyubiquitination | A cell-intrinsic factor that limits influenza virus and other viral infections such as SARS | In vitro: HEK293T, A549, NCI-H358, NCI-H2009 cells and MEFs | [56] | ||
| IGF1R (Insulin-like growth factor 1 receptor) | 🗸 | Ubiquitination | A tyrosine kinase receptor. Expression can be downregulated by NEDD4-1 through the indirect effect on the oxidisation of very-low-density lipoproteins. Ubiquitination and degradation require a C1060 site | In vitro: Hepatocytes from Landes goose embryos In vivo: Intracerebral haemorrhage mice, Sprague Dawley and Tg2576 mice |
[57] | |||
| IGPR-1 (Immunoglobulin and Proline-rich receptor-1, also known as TMIGD2/CD28H) | 🗸 | PPR | Polyubiquitination | A cell adhesion molecule involved in, for example, autophagy, angiogenesis and cell adhesion. Is degraded via the lysosomal pathway | In vitro: HEK293 cells | [58] | ||
| IRS-2 (insulin receptor substrate) | 🗸 | Monoubiquitination | NEDD4-1 recruits IRS-2 to the membrane to enhance IGF signalling | In vivo: Zebrafish embryos In vitro: HEK293 cells |
[59] | |||
| ISG15 (Interferon-stimulated gene 15) | 🗸 | ISGylation | Can attach to NEDD4-1 inhibiting its ubiquitination properties | In vitro: HEK293, HeLa cells and MEFs | [60,61,62] | |||
| KLF8 (Krueppel-like factor 8) | 🗸 | Ubiquitination | The function of this transcription factor is regulated by NEDD4-1 | In vitro: HEK293 cells | [63,64] | |||
| LATS1 (large tumour suppressor kinase 1) | 🗸 | Ubiquitination | A serine/threonine kinase involved in the suppression of tumours | In vitro: HEK293 cells | [65] | |||
| LC3 (Microtubule-associated protein 1A/1B-light chain 3) | 🗸 | WEII, WVVL, WFFL, WDKL | An autophagy-related protein. LC3 binds to NEDD4-1, but is not a ubiquitination substrate of NEDD4-1 | In vitro: HEK293 cells | [66] | |||
| LDLRAD4 (Low density lipoprotein receptor class A domain containing 4) | 🗸 | Ubiquitination | Degraded via the lysosome and is a negative regulator of TGF-β signalling | In vivo: Female BALB/c nude mice In vitro: L02 and HepG2 cells |
[67] | |||
| LMP2A (Latent membrane protein 2A) | 🗸 | PPPPY | Ubiquitination | A latent Epstein–Barr virus (EBV) infection protein involved in B cell signal transduction | In vitro: BJAB, Ramos, Raji, Jurkat, HPB.ALL and M12 cells | [68] | ||
| MDM2 (Mouse double minute 2 homolog) | 🗸 | Polyubiquitination (K63-linked) | Is an E3 ubiquitin ligase involved in negative regulation of p53. MDM2 is stabilised via NEDD4-1 interacting with its RING domain. NEDD4-1 overexpression reduces p53 levels | In vivo: NEDD4-1 KO mouse embryonic fibroblasts | [69] | |||
| MEKK5 (mitogen-activated protein kinase 5) (Apoptosis Signal-regulating Kinase 1 (ASK1) | 🗸 | A serine/threonine kinase that regulates NEDD4-1 cell migration signalling in lung cancer | In vitro: HEK293T, NCI-H1650, and A549 cells | [58] | ||||
| N4BP (NEDD4-binding protein) | 🗸 | PPLP, PPEY, PPPY | Monouniquitination and Polyubiquitination | N4BP is degraded via the proteasome. NEDD4-1 regulates N4BP1 at promyelocytic leukaemia nuclear bodies | In vitro: HEK293 cells and MEFs | [70,71] | ||
| NAB (N-aryl benzdimidazole) | 🗸? | NAB2 reduces the ratio of K63-linked ubiquitin chains on A53T α-synuclein by an unknown mechanism. NAB2 binds NEDD4-1 but does not alter conformation or enzymatic activity. | In vitro: SH-SH5Y cells | [72,73] | ||||
| NEDD4-1 (Neural precursor cell-expressed developmentally-down-regulated protein 4-1) | 🗸 | 🗸 | Autoubiquitination | K29-linked autoubiquitination. C2 and HECT domains bind resulting in autoubiquitination | In vitro: HEK293T, HeLa, THP-1 and A549 cells | [74,75] | ||
| NHE1 (Sodium-Hydrogen antiporter 1) | 🗸 | Multi-monoubiquitination and polyubiquitination | The Na(+)/H(+) exchanger 1 is ubiquitinated for degradation by NEDD4-1 but requires β-arrestin 1 | In vitro: HEK293 cells | [34] | |||
| N-Myc (N-myc proto-oncogene protein/ basic helix-loop-helix protein 37) | 🗸 | Polyubiquitination | An oncoprotein that is degraded via the proteasome. NEDD4-1 suppresses neuroblastoma and pancreatic cancer cell proliferation | In vitro: Neuroblastoma BE(2)-C, CHP134, pancreatic cancer MiaPaca-2 and HEK293 cells | [76] | |||
| Notch | 🗸 | PPSY | Polyubiquitination | A plasma membrane receptor that is ubiquitinated for degradation via the proteasome | In vivo: Drosophila and Conditional NEDD4-1 overexpression in Wistar Rat | [77,78,79] | ||
| Rap2a (RAS-related protein 2a) | 🗸 | Monoubiquitination of K63 | Rap2a Is a member of the RAS-related protein family. NEDD4-1 Inhibits GTP-Rap2a activity subsequently promoting the migration and invasiveness of glioma cells | In vitro: Human glioma cell lines U251 and U87 | [80] | |||
| RAS (Rat sarcoma virus) | 🗸 | PPGY, PPDY | Polyubiquitination and monoubiquitination | Small guanosine triphosphatases involved in a multitude of different cellular processes by acting as a molecular switch. RAS is regulated via NEDD4-1ubiquitination sending it for degradation to the lysosome. This regulation suppressed tumorigenesis | In vitro: HEK293T, HeLa, NIH 3T3, MEF and HepG2 cells | [40,81] | ||
| RET (Rearranged during transfection) | 🗸 | Polyubiquitination | A receptor tyrosine kinase. The short form (Ret9) becomes localised and internalised into the endosomal network through clathrin-coated pits following NEDD4-1 ubiquitination. This causes inhibition of Ret9-mediated neurotrophic signalling at the cell surface and promotion of post-internalisation signalling. This mechanism could potentially impact neurotrophic signalling of dopaminergic neurons and play a role in Parkinson’s disease | In vitro: HEK293 and SH-SY5Y cells | [82] | |||
| RNAPII (Ribonucleic acid Polymerase II) | 🗸 | Monouniquitination and polyubiquitination | A multiprotein involved in the transcription of DNA into mRNA that is degraded via the proteasome after being ubiquitinated by NEDD4-1. This ubiquitination is dependent on NEDD4-1 interacting with the ElonginA/B/C-Cullin 5 complex | In vitro: HEK293, MRC5 and S. cerevisiae | [83,84] | |||
| RTP801/REDD1 | 🗸 | Polyubiquitination | A pro-apoptotic protein that is targeted for degradation by NEDD4-1via K63 ubiquitin linkages. NEDD4-1 loss may elevate RTP801 proteins leading to an increase in neuronal death in Parkinson’s disease | In vivo: NEDD4-1f/f, Emx1Cre mice In vitro: PC12, HEK293 cells and rat primary cortical neurons |
[85] | |||
| SAG (S-Arrestin) | 🗸 | Polyubiquitination | An anti-apoptotic cellular survival protein that is degraded by the proteasome. NEDD4-1 reduction of SAG resulted in etoposide-induced apoptosis in cancer cells. SAG does not bind to WW domains as it lacks PY motifs but interacts with NEDD4-1 via its RING domain | In vitro: HEK293T | [86] | |||
| SCAMP3 (Secretory Carrier Membrane Protein 3) | 🗸 | PPAY, PSAP, PTEP | Multi-monoubiquitination | Integral membrane proteins involved in the cell surface recycling system. SCAMP3 is a NEDD4-1 substrate that is involved in the degradation of EGFR via the lysosome | In vitro: HeLa and HEK293T cells | [87] | ||
| Spy1A | 🗸 | Polyubiquitination | A cyclin-like protein that is needed for a cell to progress through the G1/S phase. Spy1A is required for p53-mediated tumour suppression. Spy1A is degraded in a cell cycle-dependent manner during mitosis via the ubiquitin-proteasome system | In vitro: Human mammary breast cancer, MCF7, and HEK293cells | [88,89] | |||
| SQSTM1 (p62) | 🗸 | Polyubiquitination (K63-type) | An autophagy-related protein. NEDD4-1 ubiquitinates its PB1 domain. Lack of NEDD4-1 leads to accumulation of aberrant enlarged inclusion bodies | In vitro: HEK293T, HEK293A and A549 cells | [66,90] | |||
| VEGF-R2 (vascular endothelial growth factor receptor-2) | 🗸 | Monoubiquitination | This receptor is degraded by NEDD4-1 but Grb10 regulates this process by interacting with NEDD4-1 | In vitro: HEK-293 EBNA cells | [91] |
?, experimental uncertainty. Further investigation required to confirm result. Abbreviations: PDY, Proline Aspartate Tyrosine; PDNEAYEMP, Proline Aspartate Asparagine Glutamate Alanine Tyrosine, Glutamate, Methionine Proline; PLY, Proline Leucine Tyrosine; PPLP, Proline Proline Leucine Proline; PPSP, Proline Proline Serine Proline; PFY, Proline Phenylalanine Tyrosine; LPLY, Leucine Proline Leucine Tyrosine; PPGY, Proline Proline Glycine Tyrosine; PPDY, Proline Proline Aspartate Tyrosine; PPPY, Proline Proline Proline Tyrosine; VLLVRPSRLSSSG, Valine Leucine Leucine Valine Arginine Proline Serine Arginine Leucine Serine Serine Serine Glycine; PTAP, Proline Threonine Alanine Proline; PPAY, Proline Proline Alanine Tyrosine; PPRY, Proline Proline Aspartate Tyrosine; PPEY, Proline Proline Glutamate Tyrosine; PPNY, Proline Proline Asparagine Tyrosine; PPR, polyproline rich; WEII, Tryptophan Glutamate Isoleucine Isoleucine; WVVL, Tryptophan Valine Valine Leucine; WFFL, Tryptophan Phenylalanine; WDKL, Tryptophan Aspartate Lysine Leucine; PPPPY, Proline Proline Proline Proline Tyrosine; PPSY, Proline Proline Serine Tyrosine; PSAP, Proline Serine Alanine Proline; PTEP, Proline Threonine Glutamate Proline.